US010159212B1 (12 ) United States Patent (10 ) Patent No. : US 10 , 159 ,212 B1 Simon et al. (45 ) Date of Patent: Dec . 25 , 2018 Pyne et al. , “ A first linkage map and downy mildew resistance QTL ( 54 ) DOWNY MILDEW RESISTANT/ TOLERANT discovery for sweet basil ( Ocimum basilicum ) facilitated by double SWEET BASIL VARIETIES digestion restriction site associated DNA sequencing (ddRADseq ) ," PLoS One 12 : e0184319 , 2017 . ( 71 ) Applicant: Rutgers, The State University of New Pyne et al ., “ A Rapid Screening Approch to Identify Resistance to Jersey , New Brunswick , NJ (US ) Basil Downy Mildew ( Peronospora belbahrii ), ” HortScience 49 : 1041 1045 , 2014 . Pyne et al. , “ Inheritance of Resistance to Downy Mildew in Sweet ( 72 ) Inventors : James E . Simon , Princeton , NJ (US ) ; Basil, ” J Amer Soc Hort Sci. 140 :396 -403 , 2015 . Robert Michael Pyne , Palm Desert , Pyne, “ A first linkage map and downy mildew resistance QTL CA (US ) ; Christain Andrew discovery for sweet basil ( Ocimum basilicum ) facilitated by double Wyenandt, Elmer, NJ (US ) digestion restriction site associated DNA sequencing (ddRADseq ), " PLoS One , Manuscript Draft , 2017 , 62 pages . ( 73 ) Assignee: Rutgers , The State University of New Simon et al. , “ Basil : A source of essential oils , " pp . 484 - 489 in Advances in New Crops, J . Janick and J . E . Simon ( eds . ) , Timber Jersey, New Brunswick , NJ ( US) Press , Portland , OR , 1990 . Simon et al. , “ Basil : A source of aroma compounds and a popular ( * ) Notice : Subject to any disclaimer , the term of this culinary and ornamental herb ,” pp . 499 - 505 in Perspectives on New patent is extended or adjusted under 35 Crops and New Uses , J . Janick (ed . ) , ASHS Press , Alexandria , VA , U . S . C . 154 ( b ) by 0 days. 1999 . Villani et al. , “ An improved clearing and mounting solution to replace choral hydrate in microscopic application , ” Applications in (21 ) Appl . No. : 15 / 812, 968 Plant Sciences 1 : 1300016 , 2013 . Wyenandt et al. , “ Basil Downy Mildew ( Peronospora belbahrii ) : ( 22 ) Filed : Nov . 14 , 2017 Discoveries and Challenges Relative to Its Control , ” Phytopathol. 105 :885 - 894 , 2015 . (51 ) Int . Ci. Wyenandt et al. , " Susceptibility of Basil Cultivars and Breeding AQIH 5 / 12 (2018 . 01) Lines to Downy Mildew ( Peronospora belbahrii) ,” HortTech 45 : A01H 6 /50 1416 - 1419 , 2010 . ( 2018 .01 ) Ocimum basilicum L . “ Prospera ’ , Application No . 2017 /2792 , Com ( 52 ) U . S . CI. munity Plant Variety Office Gazette , p . 61, Feb . 15 , 2018 . CPC .. .. .. A01H 6 /506 (2018 .05 ) ; A01H 5 / 12 Pyne et al ., " Population structure , genetic diversity and downy (2013 . 01 ) mildew resistance among Ocimum species germplasm ,” BMC Plant (58 ) Field of Classification Search Biol . 18 :69 , 2018 . None Ben - Naim et al . , “ Transfer of Downy Mildew Resistance from Wild Basil (Ocimum americanum ) to Sweet Basil ( O . basilicum ) , " See application file for complete search history . Phytopathology 108 : 114 - 123 , 2018 . ( 56 ) References Cited * cited by examiner U . S . PATENT DOCUMENTS Primary Examiner - David H Kruse 2018/ 0265887 AL 9 / 2018 Acosta et al . ( 74 ) Attorney , Agent, or Firm — Klarquist Sparkman , LLP OTHER PUBLICATIONS (57 ) ABSTRACT Grayer et al 1996 , Phytochemistry 43 ( 5 ) : 1033 - 1039 . * Hao et al 1996 , Phytochemistry 43 ( 4 ) : 735 -739 . * Eight new sweet basil (Ocimum basilicum L . ) cultivars Homa et al. , “ Evaluation of Fungicides for the Control of Peronospora designated ' Rutgers Thunderstruck - DMR ' , 'Rutgers Pas belbahrii on Sweet Basil in New Jersey, ” Plant Dis . 98 : 1561- 1566 , sion - DMR ' , Rutgers Obsession - DMR ' , ' Rutgers Devotion 2014 . DMR ' , “ 26 _ 24 _ 33 ”, “ 50 _ 03 _ 05 ' , “ 50 _ 03 _ 34 ', and Homa et al. , “ Morphological Characteristics and Susceptibility of * 42 _ 21 _ 02 ' are provided , as are parts of the plants , extracts Basil Species and Cultivars to Peronospora belbahrii, " HortScience and biomasses from the variety , and uses thereof, for 51 : 1389 - 1396 , 2016 . example as a food or in a food product . These plants have Koroch et al. , " Rapid staining method to detect and identify downy mildew (Peronospora belbahrii) in basil, ” Applications in Plant basil downy mildew resistance / tolerance . Sciences 1 : 1300032 , 2013 . McGrath , M . T. , Cornell University : Vegetable MD Online, “ Expect 30 Claims, 7 Drawing Sheets and Prepare for Downy Mildew in Basil ,” vegetablemdonline .path . (6 of 7 Drawing Sheet( s ) Filed in Color) cornell. edu /NewsArticles /BasilDowny .html , 2011 . Specification includes a Sequence Listing . U . S . Patent Dec . 25 , 2018 Sheet 1 of 7 US 10 , 159, 212 B1 : . : . : . : . : . .. k3.2 . k3.3 . .. : . .W W . .. XXXXX . .. k1.2 . enlang . .. 2 . A . I . k2hybrids . O .. .. + . * . k22.1xki . palingbet . ! . *** . XXX . - . kixk2 . V . FIG.1 . kl.2 U . S . Patent Dec . 25 , 2018 Sheet 2 of 7 US 10 , 159 ,212 B1 . ' ' ' ' ' ' . FIG.2B . XXX . FIG.2A . X . .' * * * . U . S . Patent Dec . 25 , 2018 Sheet 3 of 7 US 10 , 159 ,212 B1 areadtiempo . yentongangtaonnating . , ' . 4 . 9 . FIG.3 . ! U . S . Patent Dec . 25 , 2018 Sheet 4 of 7 US 10 , 159, 212 B1 * * * * * . * ** . : . : . : : : : : wwwX wwwwww . W WwwsexNASTO W x . WWW SON A 20-12 2003 . * * TORTY XXXXXX 86-9092 - 50-0334 . FIG.4A U . S . Patent Dec . 25 , 2018 Sheet 5 of 7 US 10 , 159 ,212 B1 . TIX" . * . * . * . * . 111 . 1 . 1 . 1 . XX . XXX XX . + ++ . XXXXXXXXXXXX . XXXXXXYYYYYYNXXDELITEHitit T* TIH . 111111111111111111:11144 . SITEITETITETITETIT . XXXXXX . WW . FIG.4B . 4 U . S . Patent Dec. 25 , 2018 Sheet 6 of 7 US 10 , 159, 212 B1 . :. wwwwwww . * . 2 . .. .. .. : 2 . .. areoneanother .. e *2 . wherew . * . : . .: . maniereomnisiste . *. .. .. .. .. .. *000. * . * . * . * . :. * X . la . * . * . an . n . * . w . *ytetache . tietoimin * . : FIG.5 : von 32OIL atent Dec . 25 , 2018 Sheet 7 of 7 US 10 , 159 ,212 B1 . 25-2433 . ' Y . 350-0334. Sim* -}472107 . * . ' . ' ' . V . V . .' . ' . ' . -oner. 0. .0 i . 0 . 9 . V -* . *. .* -4 . 7 . 4 . 4 . , . -. -. ' . * . .4. - . ha . *- . 24-2 . - . 44Y . N . - . .* . W . vWWW* . 4 . ' . ! . ' . .4 ' . -. ' . .444444 . W ' . ' . ! . ' . .- . *. .9 . ! . * . -. .- . +.44 . - . .- . - . - . 4. - . - - . - . V Y . Y . YYY Y Y .*,-: :*' :. FIG.6 + . US 10 , 159 ,212 B1 DOWNY MILDEW RESISTANT/ TOLERANT aroma/ flavor , the physical appearance of the plant ( pheno SWEET BASIL VARIETIES type ) varies according to its utility in the 1 ) fresh ; 2 ) processed consumer market ; or 3 ) ornamental/ home horti ACKNOWLEDGMENT OF GOVERNMENT cultural and potted plant markets . Within the processed 5 market basil is primarily consumed as a 2A ) frozen or 2B ) SUPPORT dried or freeze dried product. It is generally observed that market 1B prefers a concave leaf shape , while markets 1A , This invention was made with government support under 2A and 2B prefer a less concave ‘ large leaf' type . The large 2011 - 51181 - 30646 and 2016 -68004 - 24931 awarded by The leaf type basil varieties are associated with high yield ( i . e . , United States United Department of Agriculture (USDA ) , foliar volume) with a leaf- to.